环介导等温扩增技术快速检测食品过敏原牡蛎成分

Rapid detection of food allergen oysters by loop-mediated isothermal amplification

中文关键词:  食品过敏原  牡蛎成分  环介导等温扩增技术

英文关键词:food allergen  oysters  loop-mediated isothermal amplification

中文摘要:

      目的 建立环介导等温扩增(loop-mediated isothermal amplification, LAMP)方法快速检测食品中的过敏原牡蛎成分。方法 根据国家生物技术信息中心的牡蛎线粒体序列, 通过Primer Explorer version 5.0软件设计引物并筛选出LAMP特异性扩增引物。并进一步对反应体系优化, 对该方法的灵敏度、特异性以及稳定性进行验证。对10种牡蛎阳性样品、14种阴性样品、4类牡蛎相关食品进行检测。结果 该方法可以检测出含牡蛎成分0.1%, DNA浓度为0.01 ng/μL的样品。在实验时间上大幅缩短, 反应可在25~45 min内结束, 并且可以在微量体系下完成, 对于食品相关产品的检出率为100%。结论 该方法操作简单、成本较低、特异性高、灵敏度好, 适用于食品中过敏原牡蛎成分的检测。

英文摘要:

      Objective To establish a method for rapid detection of food allergens oyster ingredients by loop-mediated isothermal amplification (LAMP). Methods According to the mitochondrial sequences of oysters from the national biotechnology information center, primers were designed by using Primer Explorer version 5.0 software, and LAMP specific amplification primers were selected. Then the reaction system was optimized, and the sensitivity, specificity and stability of the method were verified. Totally 10 kinds of oyster positive samples, 14 kinds of negative samples and 4 kinds of oyster related food were detected. Results The method could detect the samples with 0.1% oyster composition and DNA concentration of 0.01 ng/μL. The test time was greatly shortened, could be completed within 25 to 45 min, and could be completed under a trace system, with the detection rate of 100% for food-related products. Conclusion The method is simple operation, low cost, high specificity and high sensitivity, and is suitable for detecting allergen oyster components in foods.